Localization of beta-(1,3)-glucanase in the mycelium of Sclerotium rolfsii

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Abstract
The role of the lytic enzyme .beta.-(1,3)-glucanase in cell wall synthesis and its distribution in the mycelium of the fungus S. rolfsii were studied. Enzyme activity was determined after enzyme extraction with Triton X-100 from a cell wall preparation. Specific zones of immunofluorescence appeared in the hyphal tips, clamp connections, new septa and lateral branching when a specific antiserum [rabbit] was used with the indirect method of the fluorescent antibody staining. Enzymatic activity in the cell wall preparation was inactivated by diethylpyrocarbonate. However, 69% of the total enzymatic activity was present in a latent form which was not affected by the ester. Apparently most of the .beta.-(1,3)-glucanase was present along the hyphal cell walls in a masked form. An active enzyme appeared only in those regions which showed immunofluorescence. The activity of glucan synthetase, an enzyme essential for wall formation, was higher in the branching fungus grown on L-threonine-supplemented synthetic medium than in the synthetic medium-grown fungus.